Department of Nano Engineering, Center for Nano Manufacturing, Inje University, Gimhae 621-749, Republic of Korea.
Anal Chem. 2013 Mar 5;85(5):2779-86. doi: 10.1021/ac303284u. Epub 2013 Feb 19.
This paper presents a circulating tumor cell (CTC) microseparator for isolation of CTCs from human peripheral blood using immunomagnetic nanobeads with bound antiepithelial cell adhesive molecule (EpCAM) antibodies that specifically bind to epithelial cancer cells. The isolation is performed through lateral magnetophoresis, which is induced by high-gradient magnetic separation technology, involving a ferromagnetic wire array inlaid in the bottom substrate of a microchannel. Experimental results showed that the CTC microseparator isolates about 90% of spiked CTCs in human peripheral blood at a flow rate of up to 5 mL/h and purifies to approximately 97%. The overall isolation procedure was completed within 15 min for 200 μL of peripheral blood. CTCs from peripheral blood of patients with breast and lung cancers were isolated with the CTC microseparator, and the results were compared with those of healthy donors. Using a fluorescence-based viability assay, the viability of CTCs isolated from peripheral blood of patients with cancer was observed. In addition, the usefulness of the CTC microseparator for subsequent genetic assay was confirmed by reverse-transcriptase polymerase chain reaction (RT-PCR) amplification of cancer-specific genes using CTCs isolated from patients with cancer.
本文提出了一种循环肿瘤细胞（CTC）微分离器，用于使用结合了上皮细胞黏附分子（EpCAM）抗体的免疫磁性纳米珠从人外周血中分离 CTC，EpCAM 抗体特异性结合上皮癌细胞。通过侧向磁泳，在高梯度磁分离技术的诱导下，在外周血中以高达 5mL/h 的流速分离约 90%的掺入 CTC，并将其纯化至约 97%。对于 200μL 的外周血，整个分离过程在 15 分钟内完成。使用 CTC 微分离器从乳腺癌和肺癌患者的外周血中分离 CTC，并将结果与健康供体进行比较。使用基于荧光的活力测定法，观察到从癌症患者外周血中分离的 CTC 的活力。此外，通过使用从癌症患者中分离的 CTC 进行逆转录聚合酶链反应（RT-PCR）扩增癌症特异性基因，证实了 CTC 微分离器对后续遗传检测的有用性。
